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Pig-human xenotransplantation can trigger cell-mediated immune responses. We explored the role of gangliosides in inflammation related to immune rejection in xenotransplantation. Co-culture of xenogeneic cells (pig-MSCs and RAW264.7) was used to emulate xenotransplantation conditions. MTT assay results indicated that cell viability was significantly decreased in pADMSCs co-cultured with RAW264.7 cells. GM1 and GM3 were highly expressed in pADMSCs co-cultured with RAW264.7 cells. pADMSCs co-cultured with RAW264.7 cells strongly expressed pro-inflammatory proteins such as COX-2, iNOS, p50, p65, pIκBα, and TNF-α. GM1-knockdown pADMSCs co-cultured with RAW 264.7 cells did not show significantly altered cell viability, but pro-inflammatory proteins were markedly inhibited. Co-culture of pADMSCs with RAW264.7 cells induced significant phosphorylation (p) of JNK1/2 and pERK1/2. However, pERK1/2 and pJNK1/2 were decreased and MEK1/2 and Raf1 were suppressed in GM1-knockdown pADMSCs co-cultured with RAW264.7 cells. Thus, the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways were significantly upregulated in response to increases of GM1 in co-cultured xenogeneic cells. However, the inflammatory response was suppressed in co-culture of GM1-knockdown pADMSCs with RAW264.7 cells via down-regulation of the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways. Therefore, the ganglioside GM1 appears to play a major role in the inflammatory response in xenotransplantation via the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways.

Plant-derived multiple monoclonal antibody (mAb) CO17-1A × BR55 (mAbP CO17-1A × BR55) produced in transgenic tobacco plants were cross-pollinated with mAb CO17-1A and mAb BR55. Human anti-colorectal cancer multiple mAb CO17-1A × BR55 was cloned using pBI121 vector. Mice were given a single intraperitoneal injection of azoxymethane (AOM) with 10 mg/kg body weight. Starting 1 week after the injection, mice received 2% dextran sulfate sodium (DSS) in the drinking water for 1 week. In addition, the mice were injected intraperitoneal with mAbs dissolved in phosphate buffered saline (100 μg/mouse) twice per week for 4 weeks. Apoptotic cell death, expression of pro-apoptotic proteins, activity of inflammatory cytokines and ERK pathway phosphorylation were assayed by Western blot and TUNEL kit. mAbP CO17-1A × BR55 meaningfully and efficiently suppressed the development of AOM/DSS-induced colorectal inflammation and colorectal tumors, as determined by a reduced activation of inflammatory cytokines, number ...

Gangliosides are ubiquitous components of cell membranes that can act as mediators of cell adhesion and signal transduction. However, their interaction with adhesion molecules in vascular grafts following xenotransplantation is not yet clearly understood. In this study, human umbilical vein endothelial cells (HUVECs) were cocultured with porcine aortic endothelial cells (PAECs) in order to investigate the expression patterns of gangliosides, inflammation factors, and the mRNA levels of adhesion molecules by PCR and western blot. The expression of gangliosides GM3 and GM1 was detected in both HUVECs and PAECs, while GD3 was expressed only in PAECs. However, when HUVECs were cocultured with PAECs, GD1a expression was newly detected. In addition, we observed the mRNA expression levels of adhesion molecules including such as E-selectin, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and platelet endothelial cell adhesion molecule-1 were higher in cocultured HUVECs with PAECs than in eit...

Human dental pulp-derived stem cells (hDPSCs) have been considered as alternative sources of adult stem cells because of their potential to differentiate into multiple cell lineages. Gangliosides, which sialic acid-containing glycosphingolipids are abundantly expressed in the plasma membrane and which play an important role in various processes, such as proliferation and differentiation. This study investigated the possible role of gangliosides in dopaminergic neural differentiation. When hDPSCs were cultured under dopaminergic neural differentiation conditions, the expression of dopaminergic neural cell markers genes such as TH, Pitx-3, Nurr-1, and DAT were detected. Immunofluorescence analysis showed that ganglioside biosynthesis was associated with the dopaminergic neural differentiation of hDPSCs. Specifically, GD3 and GT1b were expressed during dopaminergic neural differentiation. These results suggest that gangliosides may play a role in the dopaminergic neural differentiation process of hDPSCs.

Gangliosides are a ubiquitous component of the membranes of mammalian cells that have been suggested to play important roles in various cell functions such as cell–cell interaction, adhesion, cell differentiation, growth control and signaling. However, the role that gangliosides play in the immune rejection response in xenotransplantation is not yet clearly understood. In this study, differential expression patterns of gangliosides in HEK293 (human embryonic kidney cells), PK15 (porcine kidney cells), NIH-kd (NIH-mini pig kidney cells, primary cultured) and the cortex, medulla and calyx of the NIH-mini pig kidney were investigated by high-performance thin-layer chromatography (HPTLC). The results revealed that HEK293, PK15 and NIH-kd contained GM3, GM2 and GD3 as major gangliosides. Moreover, GM3, which are the gangliosides of NIH-kd, were expressed at higher levels than HEK293 and PK15. Especially, GT1b were expressed in HEK293 and NIH-kd but not in PK15. Finally, GM1 and GD1a were expressed in ...